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Protein Pulldown Assays to Monitor the Composition of the Bacterial RNA Degradosome.

Accepted version
Peer-reviewed

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Type

Book chapter

Change log

Authors

Hardwick, Steven W 
Luisi, Ben F 
Marques, Marilis V 

Abstract

The method of co-immunoprecipitation (co-IP or pulldown) enables the identification of proteins interacting in macromolecular assemblies, through the purification of a key protein by affinity chromatography using specific antibodies immobilized on a matrix. The advantages of using epitope-tagged proteins include the ability to use commercially available antibodies for affinity purifications, and typically they do not disrupt the structure of the protein complexes. Here we describe the utilization of an epitope-tagged version of Caulobacter crescentus RNase E in order to determine the composition of the RNA degradosome under different growth conditions. Several proteins that interact with the RNA degradosome were identified.

Description

Title

Protein Pulldown Assays to Monitor the Composition of the Bacterial RNA Degradosome.

Keywords

Affinity purification, Caulobacter, Epitope-tagged protein, Protein pulldown assay, RNA degradosome, RNase E, Bacterial Proteins, Caulobacter crescentus, Endoribonucleases, Multienzyme Complexes, Polyribonucleotide Nucleotidyltransferase, RNA Helicases, RNA, Bacterial

Is Part Of

RNA Remodeling Proteins

Book type

Publisher

Springer US

ISBN

978-1-0716-0934-7

Rights

All rights reserved
Sponsorship
Wellcome Trust (200873/Z/16/Z)